My long term goal is to understand the effects of macromolecular crowding on[unreadable] biochemical processes by acquiring atomic-level information on proteins under[unreadable] actual biological conditions. To this end, my group has pioneered in-cell NMR.[unreadable] We have shown how to assess structure, quantify dynamics, and measure[unreadable] stability under the crowded conditions found in living Escherichia coli cells using[unreadable] this powerful new technique. Now, we want to take the next step.[unreadable] With support from a Pioneer Award, we will focus on using in-cell NMR in[unreadable] eukaryotic cells to study two key proteins in neurodegenerative diseases, the[unreadable] intrinsically disordered proteins, a-synuclein and tau. These proteins are[unreadable] excellent candidates not only because of their disease relevance, but also[unreadable] because we know that macromolecular crowding has extremely large effects on[unreadable] the properties of disordered proteins.[unreadable] Our understanding of protein structure and function has grown enormously in the[unreadable] last 100 years. We have progressed from pondering what role, if any,[unreadable] polypeptides play in the cell, to unraveling, at the atomic level, the mechanisms[unreadable] of enzymes and the molecular bases of protein-protein interactions vital to[unreadable] understanding human disease.[unreadable] Our accumulating wealth of knowledge has largely come from in vitro studies[unreadable] performed under conditions far different from those found in biology. For[unreadable] example, most biochemical examinations of protein behavior are performed at[unreadable] concentrations in the ?g-to-mg-per-mL range, but the insides of cells, where most[unreadable] proteins perform their work, have protein concentrations of >300 mg per mL.[unreadable] Thus, our knowledge comes from data acquired under conditions that are far[unreadable] from physiological relevant, and theory predicts these differences can have[unreadable] extremely large effects on biophysical parameters. Moving beyond the test tube[unreadable] by performing truly in vivo studies in living eukaryotic cells by using NMR[unreadable] spectroscopy is the next frontier in protein chemistry.